Peroxidase conjugated goat anti rabbit mouse igg h l

Murine RBM39 pAb was prepared by immunizing mice with the purified protein. Anti-PRRSV nsp2 antibody was a gift from Prof. Jun Han of China Agricultural University and Rabbit anti-PRRSV-N protein pAb was purchased from YBio Technology (YB-23941R). Mouse anti-Flag/β-actin/GADPH mAb and Peroxidase-Conjugated Goat anti-Rabbit/Mouse IgG (H+L) were purchased from Yeasen Technology. Rabbit anti-phospho-JNK1+2+3 (Thr183+Tyr185) pAb was purchased from Bioss (bs-1640R); Rabbit anti-c-Jun mAb was purchased from Abways Technology (CY5290); Rabbit anti-phospho-c-Jun (Ser73) mAb was purchased from Cell Signaling Technology (3270T); Rabbit anti-HA pAb, Goat anti-Mouse IgG (H+L) Alexa Fluor 555 and Goat anti-Mouse/Rabbit IgG (H+L) FITC was purchased from Invitrogen; Mouse anti-Flag/HA-tags beads was purchased from Abmart. Phosphatase inhibitors were purchased from APE×BIO. Quick CIP were purchased from Biolabs.

The samples (100 GV oocytes) were lysed with 50 µL RIPA buffer (Sangon Biotech, China), and the lysates were separated in SDS‐polyacrylamide gel electrophoresis at 110 V for 60 min. Separated proteins were then transferred onto a nitrocellulose filter membrane at 350 mA for 60 min in ice. The membranes were blocked in PBS with 1% BSA for 1 h at room temperature and then incubated with primary antibody overnight at 4 °C. The membrane was incubated with peroxidase‐conjugated goat anti‐rabbit/mouse IgG (H+L) (Yeasen, China). After washing for another three times, SageCapture software was used for examination and quantification.